Rotea: a closed and automated instrument for efficient cell isolation, washing and conentration in cell therapy workflows

Background & Aim BACKGROUND: Processing and delivering cell and gene therapy products is of vital importance to patients and has presented numerous challenges. Rotea's counterflow centrifugation technology enables cell separation, washing, and concentration of many cell types and can be easily programmed to serve many additional workflows. The single use kit and instrument are able to capture and sustain billions of cells in a stable, fluidized bed with flexible inputs and output volumes as low as 5 mL. AIM We set out to evaluate Rotea's ability to isolate PBMCs from leukopaks, and wash and concentrate T cells, two common and vital processes in cell therapy workflows. We also investigated the input and output capabilities of the instrument. Methods, Results & Conclusion METHODS: Protocols were written using the Rotea Protocol Builder. Results from Rotea were compared to density gradient separation for PBMC isolations. T Cells for wash and concentrate protocols were expanded for 12 days in complete Optimizer media with Glutamax, L-Gluamine, HSA (Human Serum Albumin) and Il2. Before and after processing with Rotea, cells were counted and phenotyped using flow cytometry to determine the recovery and change in viability and marker expression. RESULTS Rotea performed consistent lymphocyte separation from leukapheresis products with greater than 80% recovery. Similar to manual isolation methods, cells isolated and cryopreserved using Rotea exhibit high viability and robust expansion in culture for over 14 days. Processed cells also maintain similar percentages of helper T Cells, cytotoxic T Cells and exhausted cells before and after processing. For T Cell wash and concentrate protocols, recovery was consistently above 90% and viability was virtually unchanged after processing. The distribution of CD4 and CD8 cells was also relatively unchanged. As many as 5 billion T cells were captured in a fluidized cell bed and outputs as low as 5 mL were performed without a decrease in cell recovery. CONCLUSION Rotea is a flexible, efficient system that can isolate cells and perform wash and concentrate protocols with high recovery of cells and maintenance of viability. Cells that undergo Rotea processing maintain their phenotype and readily expand in culture. Rotea has the ability to support multiple cell therapy workflows and the scalability to take a therapy from process development to manufacturing.